Effect of thrombin and bradykinin on endothelial cell mechanical properties monitored through membrane deformation

Abstract

The endothelium is closely implicated in the control and maintenance of the vascular homeostasis. The functions of endothelial cells are highly regulated by several agonists of G protein-coupled receptors (GPCR), which can mediate signals involved in morphological remodeling. Here, we evaluated the mechanical properties of human umbilical vein endothelial cells (HUVEC) in responses to two physiological agonists namely thrombin and bradykinin. We used the atomic force microscopy (AFM) technique to study changes in cell membrane stiffness and interaction between the actin cytoskeleton and the cell membrane. HUVEC stimulated with thrombin (10 nM) and bradykinin (1 microM) showed a temporal increase in their membrane stiffness from 5.0 +/- 0.1 kPa (control) to 8.2 +/- 0.4 kPa (thrombin) and 7.3 +/- 0.5 kPa (bradykinin) and in membrane tethers elongation forces from 43.9 +/- 0.9 pN (control) to 49.5 +/- 0.8 pN (thrombin) and 53.1 +/- 0.8 pN (bradykinin). These results were consistent with the reorganization of the actin cytoskeleton observed in fluorescence microscopy. This study demonstrates that these agonists induce important modifications of the cell membrane properties that can be directly linked to the reorganization and the interaction of the actin cytoskeleton near the apical side of the membrane. These changes in the mechanical properties of endothelial cells provide relevant informations in the biological and pathophysiological behaviors of endothelial cells.